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Image cytometry-based detection of aneuploidy by fluorescence in situ hybridization in suspension.
- Source :
-
Cytometry. Part A : the journal of the International Society for Analytical Cytology [Cytometry A] 2012 Sep; Vol. 81 (9), pp. 776-84. Date of Electronic Publication: 2012 Jul 26. - Publication Year :
- 2012
-
Abstract
- Cytogenetic abnormalities are important diagnostic and prognostic criteria for hematologic malignancies. Karyotyping and fluorescence in situ hybridization (FISH) are the conventional methods by which these abnormalities are detected. The sensitivity of these microscopy-based methods is limited by the abundance of the abnormal cells in the samples and therefore these analyses are commonly not applicable to minimal residual disease (MRD) stages. A flow cytometry-based imaging approach was developed to detect chromosomal abnormalities following FISH in suspension (FISH-IS), which enables the automated analysis of several log-magnitude higher number of cells compared with the microscopy-based approaches. This study demonstrates the applicability of FISH-IS for detecting numerical chromosome aberrations, establishes accuracy, and sensitivity of detection compared with conventional FISH, and feasibility to study procured clinical samples of acute myeloid leukemia (AML). Male and female healthy donor peripheral blood mononuclear cells hybridized with combinations of chromosome enumeration probes (CEP) 8, X, and Y served as models for disomy, monosomy, and trisomy. The sensitivity of detection of monosomies and trisomies amongst 20,000 analyzed cells was determined to be 1% with a high level of precision. A high correlation (R(2) = 0.99) with conventional FISH analysis was found based on the parallel analysis of diagnostic samples procured from 10 AML patients with trisomy 8 (+8). Additionally, FISH-IS analysis of samples procured at the time of clinical remission demonstrated the presence of residual +8 cells indicating that this approach may be used to detect MRD and associated chromosomal defects.<br /> (Copyright © 2012 International Society for Advancement of Cytometry.)
- Subjects :
- Algorithms
Female
Flow Cytometry
Humans
Image Processing, Computer-Assisted
In Situ Hybridization, Fluorescence
Leukemia, Myeloid, Acute genetics
Limit of Detection
Male
Models, Biological
Reproducibility of Results
Single-Cell Analysis
Aneuploidy
Leukemia, Myeloid, Acute pathology
Leukocytes, Mononuclear pathology
Subjects
Details
- Language :
- English
- ISSN :
- 1552-4930
- Volume :
- 81
- Issue :
- 9
- Database :
- MEDLINE
- Journal :
- Cytometry. Part A : the journal of the International Society for Analytical Cytology
- Publication Type :
- Academic Journal
- Accession number :
- 22837074
- Full Text :
- https://doi.org/10.1002/cyto.a.22101